Factor VII, which is involved in the clotting cascade, has proven to be a useful therapeutic agent to treat a variety of pathological conditions. Accordingly, there is an increasing need for formulations comprising activated Factor VII polypeptides that are pharmaceutically acceptable and exhibit a uniform and predetermined clinical efficacy.
The current commercially available, recombinantly-made Factor VII polypeptide composition NovoSeven® (Novo Nordisk A/S, Denmark), is, e.g., presented as a vial (about 3.0 mL container volume) containing 1.2 mg recombinant human Factor VIIa, 5.84 mg NaCl, 2.94 mg CaCl2, 2H2O, 2.64 mg GlyGly, 0.14 mg polysorbate 80, and 60.0 mg mannitol. This product is reconstituted to pH 5.5 by 2.0 mL water for injection (WFI) prior to use, thus yielding a concentration of the Factor VII polypeptide of about 0.6 mg/mL.
For therapeutic applications where administration of larger amounts (e.g. 10-20 mg) of an activated Factor VII polypeptide (e.g. rhFVIIa) is necessary, it is inconvenient to utilize a formulation like the NovoSeven® composition, because a fairly large volume (e.g. 15-30 mL) needs to be administered, typically by injection.
Thus, there is a need for liquid pharmaceutical products comprising an activated Factor VII polypeptide in a relatively high concentration.
Liquid formulations of Factor VII polypeptides are subject to degradation by autolysis. This is particularly acute for high concentration formulations and limits the liquid stability. Liquid formulations of Factor VII polypeptides containing Factor VII inhibitors/stabilizers have previously been described. However, these Factor VII inhibitors/stabilizers must be injected together with the Factor VII polypeptide molecule, and the effect of the Factor VII inhibitors/stabilizers on humans is generally not known.
WO 2005/002615 A1 discloses a liquid, aqueous pharmaceutical composition comprising a Factor VII polypeptide; a buffering agent suitable for keeping pH in the range of from about 5.0 to about 9.0; at least one metal-containing agent, wherein said metal is selected from the group consisting of first transition series metals of oxidation state +II, except zinc; and a non-ionic surfactant. Some ions of this class, e.g. cupper, have known toxicity, which can make some formulations of this type unsuitable for administration to humans.
WO 2005/016365 A1 discloses a liquid, aqueous pharmaceutical composition comprising at least 0.01 mg/mL of a Factor VII polypeptide (i); a buffering agent (ii) suitable for keeping pH in the range of from about 5.0 to about 9.0; and at least one stabilising agent (iii) comprising a —C(═N—Z1—R1)—NH—Z2—R2 motif (e.g. a benzamidine or an arginine). Some compounds of this class, e.g. L-arginine, are known to have a blood pressure lowering effect, which can make some formulations of this type unsuitable for administration to humans.
Liquid formulations are advantageous for injectable pharmaceutical products. If several injection dosages are retrieved from the same vial, a preservative is often a requirement from the regulatory authorities. A number of different compounds have been used as preservatives in injectable products (S, Nema, N. R. Washkuhn and R. J. Brendel: Excipients and their use in injectable products, PDA Journal of Pharmaceutical Science and Technology 51 (4), 166-171).
“Et studie af mikrotiterpladers anvendelse i formuleringsscreening af proteiner-med rFVIIa som modelstof” by Catharina Margrethe Lerche and Charlotte Petersen (Pharmaceutical University of Denmark, 2004) discloses formulations containing 1 mg/mL activated Factor VII and 2 mg/mL meta-cresol, but no antioxidant. These authors surprisingly found that in the presence of meta-cresol, about 30% of the protein molecules are oxidised after 4 weeks of incubation at 25° C., suggesting that a formulation with preservatives is highly detrimental to the stability of the protein.